Herbal compositions for the treatment of mucosal lesions

ABSTRACT

The present invention provides therapeutic compositions comprising extracts of the plant species  Echinacea purpurea  and  Sambucus nigra  and the extract(s) of at least one further plant selected from the group consisting of  Hypericum perforatum, Commiphora molmol  and  Centella asiatica . The compositions of the invention are of particular utility in the management of inflammatory mucosal diseases of both viral and non-viral origin.

CROSS RELATED APPLICATION

This application is a continuation of U.S. patent application Ser. No.12/941,535 filed Nov. 8, 2010, which is a divisional of U.S. patentapplication Ser. No. 12/417,111 filed Apr. 2, 2009, which is adivisional of U.S. patent application Ser. No. 10/478,718 filed Nov. 24,2003, which is a U.S. national phase of international applicationPCT/IL02/00402, filed in English on 22 May 2002, which designated theUS. PCT/IL02/00402 claims the benefit of priority to IL Application No.143318, filed 23 May 2001. The entire contents of these applications areincorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to herbal compositions useful for thetreatment of mucosal lesions. Although primarily intended for oral usethe composition may also be used on the labial, genital and othermucosal surfaces, as well as on the skin.

BACKGROUND OF THE INVENTION

Historically, the plant world has been the most important source ofmedicinal agents for the treatment of human and animal disease, and foruse as preventative agents in maintaining good health. However, for atleast the last 150 years, Western medicine has been dominated bysynthetic and/or highly purified chemical agents.

It is now being increasingly recognized, however, that plant extractsmay be highly effective agents for the prevention and treatment ofdisease. This is particularly true when one considers the low toxicityand greatly reduced incidence of adverse effects associated withplant-based medicines as compared with many synthetic or highly purifieddrugs. In addition, as the plant possesses a large number ofpharmaceutically active agents, extracts obtained therefrom exert theiractivities on a variety of physiologic processes, increasing the rangeof the desired therapeutic effect.

Although traditional reference sources of herbal medicine are valuableguides to the safe and effective use of plant extracts, the appropriateselection and combination of extracted material is still a majorchallenge to the development of new, highly effective herbal medicines.The scale of this challenge may be more clearly appreciated when it isrealized that there are approximately 750,000 species of floweringplants on earth, only very few of which have been scientifically studiedfor their potential therapeutic value.

Oral diseases constitute a diverse group of conditions that areresponsible for much human suffering. In addition to diseases of thehard tissues of the oral cavity (e.g. dental caries), there are manydifferent pathological conditions affecting the oral mucosa andperiodontal tissues. This group includes the commonly found conditionssuch as gingivitis, periodontal disease, aphthous ulceration and Herpessimplex lesions, as well as the oral manifestations of the less commonvesicular-bullous conditions such as bullous pemphigoid, pemphigus,erytheme multiforme and lichen planus, as well as other autoimmuneconditions.

The significance of host-related factors in the pathogenesis ofconditions such as periodontal disease is being increasingly recognized.Far from being a passive recipient of pathogenic agents released byplaque bacteria, the host tissues themselves (including the biochemicaland immunological factors contained therein) are now known to make anactive contribution to disease initiation and progression. One group ofhost factors which have recently received some attention in relation tothe pathogenesis of periodontal disease is the group consisting ofvarious tissue-destroying and tissue-remodelling enzymes. Of particularinterest is the large group of matrix metalloproteinases (Page, R. C.(1999) J. Periodont. Res. 34: 331-339). It is now believed that certain,defined, metalloproteinases such as matrix metalloproteinases 1-9 are ofparticular importance for the development and progression of periodontaldisease.

Although many pharmaceutical agents have been used in the management ofmucosal lesions, many of these have been relatively ineffective, whilesome (in particular, the systemic regimes) are associated withunacceptable adverse effects. There thus exists a need for new,efficacious and safe modes of treatment for many of the aforementionedmucosal diseases. There is a particular need for a safe, effectivetopical treatment.

It is a purpose of the present invention to respond to theaforementioned need by providing plant-based compositions for thetreatment of mucosal diseases.

It is another purpose of the invention to provide plant-based anti-viralcompositions for use in the treatment of oral and genital lesions.

It is a further purpose of the invention to provide compositions for thetreatment of mucosal diseases having higher efficacy and more rapidonset than compositions previously known in the art.

It is a still further purpose of the invention to provide compositionshaving lower toxicity and incidence of adverse effects thanpharmaceutical compositions for the treatment of mucosal diseases thathave been previously described in the art.

Further objects and advantages of the present invention will becomeapparent as the description proceeds.

SUMMARY OF THE INVENTION

It has now been unexpectedly found that certain compositions comprisingparticular combinations of plant extracts are highly effective in thetreatment of certain mucosal lesions, particularly those of the oral,anal and genital mucosa, as well as in the treatment of certain skinlesions. It is to be noted that the compositions, medicaments andtreatment methods of the present invention which will presently bedisclosed, described and exemplified, have been unexpectedly found tocause a dramatic improvement in two significant clinical parametersassociated with the mucosal and skin lesions being treated thereby.Firstly, it has been surprisingly found that said compositions,medicaments and treatment methods lead to unexpectedly rapid resolutionof the mucosal and skin lesions that are being treated. Secondly, thecompositions, medicaments and treatment methods of the present inventionhave also been surprisingly found to cause a dramatic reduction of thepain associated with the mucosal and skin lesions being treated thereby.

The present invention is primarily directed to a therapeutic compositioncomprising extracts of the plant species Echinacea purpurea and Sambucusnigra and the extract(s) of at least one further plant selected from thegroup consisting of Hypericum perforatum, Commiphora molmol and Centellaasiatica.

In one preferred embodiment of the therapeutic composition of thepresent invention, the extract(s) of the at least one further plant areextracts of the plant species Hypericum perforatum and Commiphoramolmol.

While it is not intended that the use of the composition of theabovementioned preferred embodiment of the composition of the inventionbe bound to, or limited by any particular theory regarding its chemicalor pharmacological mode of action, the present invention is particulardirected to an anti-viral composition comprising extracts of the plantspecies Echinacea purpurea, Sambucus nigra, Hypericum perforatum andCommiphora molmol.

In a preferred embodiment of the invention, the above-mentionedanti-viral composition further comprises extracts of plants selectedfrom the group consisting of Uncaria tomentosa, Thymus vulgaris,Matricaria recutita, Salix alba, Calendula officinalis, Usnea barbata,Ligusticum porterii-osha, Gaultheria procumbens, Camellia sinensis,Vaccinium myrtillus, Melissa officinalis, Allium sativum, Camelliasinensis and Krameria triandra.

In a particularly preferred embodiment of the invention, theabove-mentioned anti-viral composition comprises extracts of the plantspecies Echinacea purpurea, Sambucus nigra, Hypericum perforatum,Commiphora molmol and Uncaria tomentosa.

The present invention also provides a therapeutic composition comprisingextracts of the plant species Echinacea purpurea and Sambucus nigratogether with an extract of the plant species Centella asiatica.

In a preferred embodiment of the invention, the immediately precedingtherapeutic composition is intended for use in the treatment of diseasesof the oral mucosa. In a more preferred embodiment of the invention,said therapeutic composition is intended for use in the treatment of anoral mucosal disease selected from the group consisting of periodontaldisease, gingivitis, aphthous ulceration, mechanical trauma, thermaltrauma, lichen planus, bullous pemphigoid, pemphigus vulgaris,dermatitis herpetiformis, angular chelitis and recurrent herpes.

In a further preferred embodiment, the above therapeutic composition isintended for use in the treatment of skin lesions. In one preferredembodiment of the invention, said therapeutic composition is intendedfor use in the treatment of dermal trauma. In another preferredembodiment, the therapeutic composition is intended for use in thetreatment of insect bites and other local, superficial irritations.

In a still further preferred embodiment of the invention, the abovetherapeutic composition is intended for use in the treatment of anallesions. In a more preferred embodiment of the invention, saidtherapeutic composition is intended for use in the treatment of an anallesion associated with a condition selected from the group consisting ofanal fissures, hemorrhoids and non-specific irritation.

While it is not intended that the mechanism of action of the therapeuticcomposition for treating mucosal diseases that is disclosed immediatelyhereinabove be bound to any particular pharmacological orpathophysiological mechanism or mechanisms, it is believed that saidcomposition exerts at least some of its desired effects by inhibitingone or more matrix metalloproteinase (MMP) enzymes that are present inthe oral mucosal and periodontal tissues, and/or by increasing collagenproduction at or close to the mucosal site to which said composition isapplied. In particular, it is believed that said therapeuticcompositions may exert at least some of their desired effects by thespecific inhibition of certain specific enzymes of the MMP group. Morespecifically, it is believed that the therapeutic compositions of thepresent invention are specific inhibitors of MMP subclasses 1-9, stillmore specifically of subclasses 1, 2, 8 and 9.

Thus, the invention is also directed to a therapeutic compositioncomprising extracts of the plant species Echinacea purpurea, Sambucusnigra and Centella asiatica described hereinabove, for the inhibition ofone or more matrix metalloproteinases.

It is to be noted that the term “inhibition of one or more matrixmetalloproteinases” as used immediately hereinabove and hereinabove isintended to convey the meaning of the inhibition of the activity ofthese enzymes on their substrates.

In a preferred embodiment of this aspect of the invention, the one ormore matrix metalloproteinases to be inhibited are selected from thegroup consisting of matrix metalloproteinases 1-9. In a more preferredembodiment, said one or more matrix metalloproteinases are selected fromthe group consisting matrix metalloproteinases 1, 2, 8 and 9. Still morepreferably, the matrix metalloproteinase to be inhibited is matrixmetalloproteinase 2. In a still further preferred embodiment of thisaspect of the present invention, the matrix metalloproteinase-inhibitingtherapeutic compositions described immediately hereinabove are intendedfor use in the treatment of a disease of the oral mucosa selected fromthe group consisting of periodontal disease and aphthous ulceration.

In a further preferred embodiment of the invention, the aforementionedtherapeutic compositions for treating conditions of the oral or analmucosal tissues, as well as the aforementioned therapeutic compositionsfor inhibiting matrix metalloproteinases further comprise extracts ofplants selected from the group consisting of Uncaria tomentosa, Thymusvulgaris, Matricaria recutita, Salix alba, Calendula officinalis, Usneabarbata, Ligusticum porterii-osha, Gaultheria procumbens, Camelliasinensis, Vaccinium myrtillus, Melissa officinalis, Allium sativum,Camellia sinensis and Krameria triandra.

In another aspect, the present invention is directed to the use of acombination of extracts of the plant species Echinacea purpurea andSambucus nigra and of at least one further plant species selected fromthe group consisting of Hypericum perforatum, Commiphora molmol andCentella asiatica in the preparation of a medicament.

In one preferred embodiment, the invention is directed to the use of thecombination of plant extracts described immediately hereinabove in thepreparation of a medicament, wherein said extracts of at least onefurther plant are extracts of Hypericum perforatum and Commiphoramolmol. Preferably, this combination of plant extracts is used in thepreparation of an anti-viral medicament.

In a further preferred embodiment, the present invention is directed tothe use of extracts of plants selected from the group consisting ofUncaria tomentosa, Thymus vulgaris, Matricaria recutita, Salix alba,Calendula officinalis, Usnea barbata, Ligusticum porterii-osha,Gaultheria procumbens, Camellia sinensis, Vaccinium myrtillus, Melissaofficinalis, Allium sativum, Camellia sinensis and Krameria triandra, inaddition to the extracts mentioned hereinabove, in the preparation of ananti-viral medicament.

In a particularly preferred embodiment the present invention is directedto the use of a combination of extracts of the plant species Echinaceapurpurea, Sambucus nigra, Hypericum perforatum, Commiphora molmol andUncaria tomentosa in the preparation of an anti-viral medicament.

The present invention also provides for the use of the above combinationof plant extracts in the preparation of a medicament, said extract of atleast one further plant being an extract of Centella asiatica.Preferably, this combination of plant extracts is used in thepreparation of a medicament for the treatment of a disease of the oralmucosa. In one embodiment of the invention, said disease of the oralmucosa is selected from the group consisting of periodontal disease,gingivitis, aphthous ulceration, mechanical trauma, thermal trauma,lichen planus, bullous pemphigoid, pemphigus vulgaris, dermatitisherpetiformis, angular chelitis and recurrent herpes.

In another preferred embodiment, the invention also provides for the useof the above combination of plant extracts in the preparation of amedicament for the treatment of a skin lesion. In one preferredembodiment, the skin lesion to be treated is a lesion arising fromdermal trauma. In a further preferred embodiment, the skin lesion to betreated is an insect bite.

In another preferred embodiment of the invention, the abovementionedcombination of plant extracts is used in the preparation of a medicamentfor the treatment of a disease of the anal mucosa. In one embodiment ofthe invention, said disease of the anal mucosa is selected from thegroup consisting of anal fissures, hemorrhoids and non-specificirritation.

In another aspect the invention provides for the use of a combination ofextracts of the plant species Echinacea purpurea, Sambucus nigra andCentella asiatica in the preparation of a medicament for inhibiting oneor more matrix metalloproteinases. Preferably, said matrixmetalloproteinases are selected from the group consisting of matrixmetalloproteinases 1 to 9. Most preferably, the one or more matrixmetalloproteinases to be inhibited are selected from the groupconsisting of matrix metalloproteinases 1, 2, 8 and 9. In a preferredembodiment, the abovementioned metalloproteinase-inhibiting medicamentis used to treat a disease of the oral mucosa selected from the groupconsisting of periodontal disease and aphthous ulceration.

In a further preferred embodiment, the invention is directed to the useof the above combination of plant extracts in combination with furtherextracts of plants selected from the group consisting of Uncariatomentosa, Thymus vulgaris, Matricaria recutita, Salix alba, Calendulaofficinalis, Usnea barbata, Ligusticum porterii-osha, Gaultheriaprocumbens, Camellia sinensis, Vaccinium myrtillus, Melissa officinalis,Allium sativum, Camellia sinensis and Krameria triandra, in thepreparation of medicaments for the treatment of a diseases of the oraland/or anal mucosal tissues, and in the preparation of medicaments forinhibiting the abovementioned one or more matrix metalloproteinases.

In another aspect, the present invention is directed to a combination ofextracts of the plant species Echinacea purpurea and Sambucus nigra andof at least one further plant species selected from the group consistingof Hypericum perforatum, Commiphora molmol and Centella asiatica for useas a medicament.

In a preferred embodiment, the invention is directed to a combination ofextracts as disclosed immediately hereinabove, wherein the extracts ofthe at least one further plant are extracts of Hypericum perforatum andCommiphora molmol. In a preferred embodiment, the invention is directedto said combination of extracts for use as an anti-viral medicament. Ina further preferred embodiment, said combination of extracts is furthersupplemented by extracts of one or more plants selected from the groupconsisting of Uncaria tomentosa, Thymus vulgaris, Matricaria recutita,Salix alba, Calendula officinalis, Usnea barbata, Ligusticumporterii-osha, Gaultheria procumbens, Camellia sinensis, Vacciniummyrtillus, Melissa officinalis, Allium sativum, Camellia sinensis andKrameria triandra.

In a particularly preferred embodiment, the invention is directed to acombination of extracts of the plant species Echinacea purpurea,Sambucus nigra, Hypericum perforatum, Commiphora molmol and Uncariatomentosa for use as an anti-viral medicament.

The invention also provides a combination of plant extracts as disclosedhereinabove for use as a medicament, said extract of the at least onefurther plant being an extract of Centella asiatica. Preferably, thiscombination of extracts is provided for use as a medicament for thetreatment of diseases of the oral mucosa. While said combination ofplant extracts may be used as a medicament for the treatment of manydifferent conditions of the oral mucosa, in a preferred embodiment, thedisease to be treated is selected from the group consisting ofperiodontal disease, gingivitis, aphthous ulceration, mechanical trauma,thermal trauma, lichen planus, bullous pemphigoid, pemphigus vulgaris,dermatitis herpetiformis, angular chelitis and recurrent herpes. Inanother preferred embodiment, the combination of plant extracts isprovided for use as a medicament for the treatment of skin lesions. Inone preferred embodiment, the skin lesions are lesions arising fromdermal trauma. In another preferred embodiment, the skin lesions areinsect bites. In yet another preferred embodiment, the aforementionedcombination of extracts is provided for use as a medicament for thetreatment of diseases of the anal mucosa. While said combination ofplant extracts may be used as a medicament for the treatment of manydifferent conditions of the anal mucosa, in a preferred embodiment, thedisease to be treated is selected from the group consisting of analfissures, hemorrhoids and non-specific irritation.

In another aspect, the above-described combination of extracts is usedas a medicament for inhibiting one or more matrix metalloproteinases.Preferably, the one or more matrix metalloproteinases are selected fromthe group consisting of matrix metalloproteinases 1 to 9. Morepreferably, said metalloproteinases are selected from the groupconsisting of matrix metalloproteinases 1, 2, 8 and 9. In a preferredembodiment, the abovementioned combination of extracts for inhibitingmetalloproteinases is used in the treatment of a disease of the oralmucosa selected from the group consisting of periodontal disease andaphthous ulceration.

In yet another embodiment of the invention, the plant extracts used inthe aforementioned combination of extracts are further supplemented byextracts of plants selected from the group consisting of Uncariatomentosa, Thymus vulgaris, Matricaria recutita, Salix alba, Calendulaofficinalis, Usnea barbata, Ligusticum porterii-osha, Gaultheriaprocumbens, Camellia sinensis, Vaccinium myrtillus, Melissa officinalis,Allium sativum, Camellia sinensis and Krameria triandra.

The present invention also encompasses a method of treatment of mucosaland/or skin lesions comprising the application of atherapeutically-effective amount of a mixture of extracts of the plantspecies Echinacea purpurea and Sambucus nigra and the extract(s) of atleast one further plant selected from the group consisting of Hypericumperforatum, Commiphora molmol and Centella asiatica to the mucosallesions and surrounding tissue of a subject in need of such treatment.In a preferred embodiment of this method of treatment, said extracts ofat least one further plant are extracts of Hypericum perforatum andCommiphora molmol. In a preferred embodiment, the lesions to be treatedby this method of treatment are viral lesions.

In a further preferred embodiment, the present invention also provides amethod of treatment of viral lesions as described hereinabove, whereinthe aforementioned plant extracts are supplemented by extracts of plantsselected from the group consisting of Uncaria tomentosa, Thymusvulgaris, Matricaria recutita, Salix alba, Calendula officinalis, Usneabarbata, Ligusticum porterii-osha, Gaultheria procumbens, Camelliasinensis, Vaccinium myrtillus, Melissa officinalis, Allium sativum,Camellia sinensis and Krameria triandra.

In a particularly preferred embodiment, the present invention provides amethod of treatment of mucosal and/or skin lesions of viral origincomprising the application of a therapeutically-effective amount of amixture of extracts of the plant species Echinacea purpurea, Sambucusnigra, Hypericum perforatum, Commiphora molmol and Uncaria tomentosa.

In another preferred embodiment of the method of the invention, theextract of the at least one further plant is an extract of Centellaasiatica. In one preferred embodiment of this aspect of the invention,the lesions to be treated are oral lesions associated with a diseaseselected from the group consisting of periodontal disease, gingivitis,aphthous ulceration, mechanical trauma, thermal trauma, lichen planus,bullous pemphigoid, pemphigus vulgaris, dermatitis herpetiformis,angular chelitis and recurrent herpes. In another preferred embodiment,the lesions to be treated are skin lesions. In one more preferredembodiment, the skin lesions to be treated are lesions arising fromdermal trauma. In a further preferred embodiment, the lesions are insectbites. In another preferred embodiment of this aspect of the invention,the lesions to be treated are anal lesions associated with a diseaseselected from the group consisting of anal fissures, hemorrhoids andnon-specific irritation.

In a further aspect, the present invention is directed to a method ofinhibiting one or more matrix metalloproteinases in mucosal and/or skinlesions of a subject in need of such treatment, comprising theapplication of a therapeutically-effective amount of a mixture ofextracts of the plant species Echinacea purpurea, Sambucus nigra andCentella asiatica to said mucosal and/or skin lesions and surroundingtissue. Preferably, the one or more matrix metalloproteinases areselected from the group consisting of matrix metalloproteinases 1 to 9.More preferably, the one or more matrix metalloproteinases are selectedfrom the group consisting of matrix metalloproteinases 1, 2, 8 and 9. Ina preferred embodiment of this aspect of the invention, theaforementioned inhibition of the one or more matrix metalloproteinasesis used in the treatment of periodontal disease. In another preferredembodiment, the inhibition of the one or more matrix metalloproteinasesis used in the treatment of aphthous ulceration.

In each of the above-described methods, the mixture of plant extractsused may further comprise extracts of plants selected from the groupconsisting of Gotu kola, Uncaria tomentosa, Thymus vulgaris, Matricariarecutita, Salix alba, Calendula officinalis, Usnea barbata, Ligusticumporterii-osha, Gaultheria procumbens, Camellia sinensis, Vacciniummyrtillus, Melissa officinalis, Allium sativum, Camellia sinensis andKrameria triandra.

In one preferred embodiment of the invention, the anti-viralcompositions, medicaments and treatment methods are used in thetreatment or management of viral lesions of the oral cavity.

In another preferred embodiment of the invention, the anti-viralcompositions, medicaments and treatment methods are used in thetreatment or management of perioral lesions of viral origin.

In yet another preferred embodiment of the invention, the anti-viralcompositions, medicaments and treatment methods are used in thetreatment or management of genital lesions of viral origin.

In yet another preferred embodiment of the invention, the anti-viralcompositions, medicaments and treatment methods are used in thetreatment or management of viral lesions caused by the Herpes simplexvirus.

In yet another preferred embodiment of the invention, the anti-viralcompositions, medicaments and treatment methods are used in thetreatment or management of viral lesions of the skin.

In another aspect, the present invention also encompasses a method forinhibiting one or more matrix metalloproteinases in vitro, comprisingcontacting an effective amount of a mixture of extracts of the plantspecies Echinacea purpurea, Sambucus nigra and Centella asiatica withsaid one or more matrix metalloproteinases. In one embodiment of thisaspect of the invention, the one or more matrix metalloproteinases to beinhibited are selected from the group consisting of matrixmetalloproteinases 1 to 9. In another embodiment, the one or more matrixmetalloproteinases to be inhibited are selected from the groupconsisting of matrix metalloproteinases 1, 2, 8 and 9.

All the above and other characteristics and advantages of the presentinvention will be further understood from the following illustrative andnon-limitative examples of preferred embodiments thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 graphically illustrates the reduction in ulcer-associated painfollowing treatment with an herbal composition of the invention.

FIG. 2 shows a typical gelatin zymogram indicating the inhibitoryeffects of composition of the invention on the activity of a mixture ofmatrix metalloproteinases.

FIG. 3 graphically illustrates the reduction in insect bite-associatedpain and irritation following treatment with an herbal composition ofthe invention.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

The compositions and medicaments of the present invention are based onmixtures of plant extracts. It is to be noted that the term “extract” isused herein to include all of the many types of preparations containingsome or all of the active ingredients found in the relevant plants. Thusthe extracts may be produced by cold extraction techniques using avariety of different extraction solvents including, but not limited to,water, fatty solvents (such as olive oil), and alcoholic solvents (e.g.70% ethanol). Cold extraction techniques are usefully applied to softerparts of the plant such as leaves and flowers, or in cases wherein thedesired active components of the plant are heat labile. Alternatively,the aforementioned solvents may be used to produce extracts of thedesired plants by a hot extraction technique, wherein said solvents areheated to a high temperature, the precise value of said temperaturebeing dependent on the properties of the chosen solvent, and maintainedat that temperature throughout the extraction process. Hot extractiontechniques are more commonly applied to the harder, tougher parts of theplant, such as bark, woody branches and larger roots. In some cases,sequential extractions need to be performed in more than one solvent,and at different temperatures.

Standard procedures for producing plant extracts (including hotextraction, cold extraction and other techniques) are described in manypublications including “Medicinal plants: a field guide to the medicinalplants of the Land of Israel (in Hebrew), author: N. Krispil, Har Gilo,Israel, 1986” and “Making plant medicine, author: R. Cech, pub. byHorizon Herbs, 2000”.

Compositions and medicaments containing mixtures of extracts ofdifferent plant species, such as those of the present invention may beprepared using different ratios of each extract. For example, theantiviral medicaments and compositions of the present inventionpreferably comprise extracts of Echinacea purpurea, Sambucus nigra,Commiphora molmol and Hypericum perforatum in the following range ofweight ratios:

-   -   2-6:2-4:3-6:2-6

More preferably, these components are present in the weight ratio of4:3:5:4.

Similarly, the compositions of the invention used to treat mucosaldiseases preferably comprise extracts of Centella asiatica, Echinaceapurpurea and Sambucus nigra in the following range of weight ratios:

0.5-3:0.5-3:2:15

More preferably, these extracts are present in the weight ratio of1.5:1.5:7

In order to treat a patient with a therapeutic composition or medicamentcontaining a mixture of herbal extracts as described hereinabove, it isnecessary to administer said composition or said medicament in atherapeutically-effective amount, that is, in an amount that willprovide a concentration of the herbal extracts at the treatment sitethat is capable of exerting the desired therapeutic effect. It has beenfound, in general terms, that the compositions and medicaments of thepresent invention need to be administered in amounts such that,typically, each topical dose contains between 0.1 mg and 10 mg (dryweight) of each herbal extract, the precise values depending on theparticular combination of extracts used, and on the mode of topicaldelivery. Thus, in the case of the therapeutic composition of thepresent invention that is used in the treatment of mucosal lesions, theweights of the original plant material used to prepare acontrolled-release delivery device (as described in Example 2,hereinbelow) are:

Centella asiatica 1.6 mg

Echinacea purpurea 1.6 mg

Sambucus nigra 7.56 mg

In the case of compositions and medicaments intended primarily fortopical use (such as those of the present invention), it is necessary toadminister said compositions and medicaments for periods of time thatare sufficient to allow optimal contact of the therapeutically effectiveamounts of the herbal extracts with the lesions to be treated. When thecompositions and medicaments are to be given by incorporation into acontrolled-release intra-oral device (as described in Example 2hereinbelow), said device needs to remain in contact with the lesion tobe treated for a period of between 1 and 5 hours. This treatment may berepeated up to 5 times each day, as required, and as determined by acompetent clinician.

Mouthwashes containing the compositions and medicaments of the presentinvention should be taken in quantities of between 5 ml and 15 ml andallowed to remain in contact with the lesions to be treated for periodsof between 30 seconds and one minute. This treatment regime may berepeated up to 5 times per day.

Lozenges, pastilles, candies and other solid, soluble formulations areto be placed in the mouth, if possible in close proximity to the lesionsto be treated, and allowed to dissolve at the natural rate determined bythe additives present in said formulations.

The compositions and medicaments of the present invention as disclosedhereinabove and exemplified hereinabove may be prepared and delivered ina number of different forms.

In a preferred embodiment of the invention, medicaments and compositionsare intended for topical application at the site of the mucosal lesion.Dosage forms suitable for topical application to mucosal surfacesinclude ointments, pastes, lotions, creams, mouthwashes, lozenges,candies, chewing gums, solutions, gels and sprays. Thus, in addition tothe active ingredients, the compositions of the present invention mayalso contain excipients such as zinc, zinc oxide, silicones, calciumsilicate, aluminum hydroxide, polyethylene glycols, fats of animal orvegetable origin, oils, waxes gums, starch and cellulose or cellulosederivatives.

In other embodiments of the invention, compositions for vaginaladministration or for anal administration may be prepared by mixing theactive plant-derived components with suitable non-toxic, non-irritatingcarriers such as suppository wax, polyethylene glycol or cocoa butter.

In a preferred embodiment of the invention, the compositions andmedicaments are administered by means of a localized delivery systemthat allows topical release of the active constituents of saidcompositions and medicaments. Any suitable local delivery device may beused to administer the compositions and medicaments to the mucosalsurface. However, in a particularly preferred embodiment of theinvention the local delivery device is a slow release device such asillustrated hereinbelow in Example 2.

The following examples are provided for illustrative purposes and inorder to more particularly explain and describe the present invention.The present invention, however, is not limited to the particularembodiments disclosed in the examples.

Example 1 Effect of the Anti-Viral Composition on the Formation of ViralPlaques In Vitro

Method:

The antiviral composition was prepared as follows: 2 ml of a 1:1hydroalcoholic extract of Echinacea purpurea was mixed with 7.5 ml of a1:5 hydroalcoholic extract of Sambucus nigra, 8 ml of a 1:4hydroalcoholic extract of Commiphora molmol, 10 ml of a 1:4 preparationof a hydroalcoholic extract of Uncaria tomentosa and 20 ml of a 1:10hydroalcoholic extract of Hypericum perforatum. The term “a 1:xhydroalcoholic extract” as used herein indicates that 1 gram of plantmaterial was extracted with x volumes of the alcoholic extractionmedium. In the case of all of the plant extracts used in the presentexample, the extraction medium was a “hydroalcohol”. For the presentpurposes, the term “hydroalcohol” is defined as an aqueous solution of alower alcohol. Preferably, the lower alcohol used was ethanol, which wasgenerally prepared as a 50% solution. In some preparations, ethanol wasprepared at a different aqueous dilution within the range of 25-90%(v/v), with respect to the ethanol. The weight ratio of E. purpurea:S.nigra:C. molmol:U. tomentosa:H. perforatum in this mixture is 4:3:4:5:4.The above-mentioned alcoholic extracts were purchased either from HerbalApothecary, Syston, Leicester, U.K. or from Analit Extracts Ltd, M.P.Hefer 38100, Israel.

Disks of 3 MM filter paper (Whatman Inc.) (5 mm diameter) were soaked ina solution of the compositions to be tested, and placed on a semi-solidagar-containing culture medium covering a monolayer of BSC-1 (greenmonkey kidney) cells infected with a partially confluent dose of eitherHerpes simplex type 1 virus (HSV-1) or Herpes simplex type 2 virus(HSV-2). Following 3-4 days incubation at 37° C., the cells were fixedwith formaldehyde (20% aqueous solution) and stained with crystal violet(0.1% solution in 0.1M citric acid). The presence of a white color inthe central area of the culture indicated toxic damage of the culturedcells due to the anti-viral compositions. Inhibition of viral plaqueformation indicated that the composition tested possesses anti-viralactivity.

Acyclovir (ACG), a known and commonly used drug against herpes viruses,was included in the assay as a positive control.

Results:

The results of a typical plaque assay are given below.

Extract Toxicity anti-HSV1 anti-HSV2 Virosyn 0-10  2-11 3-11 Hypericum 54-7 3-12 Uncaria 0 0-8 7-8  Note 1: Virosyn is the herbal compositiondescribed hereinabove comprising extracts of the following five plantspecies: Echinacea purpurea, Hypericum perforatum, Commiphora molmol,Uncaria tomentosa and Sambucus nigra. Note 2: The numerical results inthe above table are the diameters of the plaques (in mm) after treatmentof the cell cultures with the disks soaked with extracts. Each virusinhibition or cell toxicity experiment was performed in triplicate. Note3: The toxicity of each extract was assessed by measuring the diameterof the blue-stained plaque in the center of cell cultures that did notreceive virus.

The above results indicate that the herbal extract mixture testedpossess antiviral activity for both HSV1 and HSV2 with minimal toxicityto the cultured mammalian cells.

Example 2 Topical Slow-Release Device for Delivery of the Compositionsto the Oral Mucosa

This example demonstrates the preparation of a slow-release device andthe incorporation therein of a plant extract mixture containingCentella, Echinacea and Sambucus.

The slow release device consists of a mixture of carbomer (carbopol),hydroxypropyl cellulose and magnesium stearate blended as describedhereinbelow. Magnesium stearate is used as a protective coating toreduce the solubility and adhesiveness of the device.

The device is prepared as follows:

1. The plant extract mixture is prepared by mixing 6 ml of a 1:4hydroalcoholic extract of Centella asiatica with 1.5 ml of a 1:1hydroalcoholic extract of Echinacea purpurea and 35 ml of a 1:5hydroalcoholic extract of Sambucus nigra. The weight ratio of C.asiatica:E. purpurea:S. nigra in this mixture is 15:15:70. Thehydroalcoholic extracts of Centella asiatica and Echinacea purpurea werepurchased from Herbal Apothecary, Syston, Leicester, U.K., while thehydroalcoholic extract of Sambucus nigra was purchased from AnalitExtracts Ltd., M.P. Hefer 38100, Israel. It is be noted that theabovementioned extract values of the form 1:x indicate that 1 g of theplant material was dissolved in x liters of solvent. The term“hydroalcoholic extract” indicates that the plant material was extractedusing ethanol at concentrations of between 25% and 50% in water.

2. The plant extract mixture is mixed with 2 g sucrose and evaporated todryness at 40° C. The residue is dissolved in 2 ml water, a furthersmall volume of water added, and the solution lyophilized overnight.

3. A mixture of the carbomer compound Carbopol® 934 P (B.F. Goodrich,Cleveland, Ohio, USA) (2 g) and hydroxypropyl cellulose (Klucel Type HF,Hercules BV, Rijswijk, Holland) (1 g) is prepared by crushing the twocomponents together.

4. A 1 g aliquot of the carbomer-hydroxypropyl cellulose mix (preparedin step 3.) is mixed together with 100 mg of the lyophilized plantextract powder (prepared in step 2).

5. Magnesium stearate (pharmaceutical grade, obtained from Riedel-DeHaen, Germany) (1 g) is mechanically mixed with 2 g of thecarbomer-hydroxypropyl cellulose mix.

6. 14 mg of the magnesium stearate-polymer mix (step 5.) is placed onthe bottom of the plunger (13 mm diameter die manufactured byPerkin-Elmer, U.K.) of a mechanical press (Spex Industries, Mutuchen,N.J., USA) and overlaid with 70 mg of the plant extract-polymer mix(step 4.). Pressure (10 tons force) is applied for 30 seconds.

In addition to the active herbal ingredients, various flavorings,excipients and colorings may be added in order to modify the taste,consistency and color of the preparation.

The side of the device containing the herbal ingredients (i.e. the sidenot containing the magnesium stearate) is applied directly to themucosal surface containing the lesion to be treated. Alternatively, themucosal surface may be pre-moistened with water or saline beforeapplication of the device. Following application, the device is held inplace with gentle pressure for approximately 10 seconds. After releasingthe gentle pressure, the device adheres to the mucosal tissue for aperiod of up to five hours.

Depending on the mucosal lesion to be treated, the device containing theherbal mixture described hereinabove may be used several times per day(e.g. 3 times per day) for periods of between two days and one month.

Example 3 Use of an Herbal Composition of the Invention to Reduce thePain Associated with Oral Mucosal Lesions

A convenient, non-random sample of 57 dental patients presenting in aprivate dental clinic with painful oral ulcers of either traumatic oraphthous origin were treated by applying to the affected site aslow-release device containing a herbal composition of the invention (asdescribed in Example 2 hereinabove). The device was left in place for a24 hour period. The ulcer-associated pain experienced by the patientswas recorded and expressed on a visual analog scale (S. Chrubasik et al.(2000) Am. J. Med. 109: 9-14), as depicted in FIG. 1. The clinicalcorrelates of the pain index values used in this scale are as follows:0=no pain; 50=requires analgesic; 100=requires anesthetic. The highestrecorded pain index reported by an individual patient in this study was90.

It may be seen from these results that the patients experienced analmost immediate decrease in pain (with a mean decrease of greater than50%). This decrease in pain levels continued over the following 6 hours,achieving a mean pain decrease of greater than 70%. The painful symptomsdid not recur following cessation of treatment.

Example 4 Effect of an Herbal Composition of the Invention on the Sizeof Mucosal Lesions

Operating as in the study presented in Example 3, the effect of theherbal composition used therein on the healing of the oral ulcerationexperienced by the patients was determined by quantification of lesionsize using a Scion image analysis system. Briefly, lesions werephotographed and digitized using a digital camera and associatedSmartcard. The image files obtained thereby were processed using thePhotoshop software package (Adobe Systems Inc.) running in MicrosoftWindows ME on an IBM-compatible personal computer. The periphery of eachlesion was outlined and copied into a new window of the NIH Imagesoftware package (National Institutes of Health, Bethesda, Md.), where,following thresholding, the lesion area was automatically calculated.

Results from a sequential study of 45 patients with oral ulcerationdemonstrate that the treatment with the herbal composition caused a mean60% decrease in lesional size over a 24-36 hour period.

Example 5 Anticollagenase Activity of an Herbal Composition of theInvention

Anticollagenase Testing:

Procedure:

Protease activity was assessed on gelatin zymograms. Twelve percentpolyacrylamide gels (0.75 mm thickness) were cast containing 10% gelatinas a substrate for the collagenase enzymes, which were applied to thegels under non-reducing conditions without heating. The gels were run,soaked in 200 ml of 2% Triton X-100 in distilled water on a gyratoryshaker (0.5 hours, 20° C.), and incubated in developing buffer (50 mMTris [pH 8.0], 1 mM CaCl₂), unless otherwise indicated, for 15 hours at37° C. The gels were examined following staining with Coumassie blue.Protease activity shows up as clear bands (indicative of cleavage of thegelatin substrate) on a blue background. For inhibition studies, eitherspecific protease inhibitors (DFP (1 Mm), EDTA (5 Mm), BBI (10 mg/ml),phenylmethyl sulfonyl fluoride (PMFS) (50 Mm) or tetracyclines (0.1 and0.25 Mm)) or a composition comprising a mixture of Echinacea purpurea,Sambucus nigra and Centella asiatica (prepared as described in Example2, hereinabove) were added to the developing buffer after the run butbefore the gel was incubated in said developing buffer. In the lattercase, the herbal composition was added to the buffer at a concentrationof one volume composition to 50 volumes buffer. To determine proteaseactivity as a function of pH, samples were run on zymograms andsubsequently incubated in the appropriate buffer (50 Mmcitrate-phosphate buffer [pH 5], 50 Mm ADA buffer [pH 6 and 7], 50 MmTRIS [pH 8 and 9] or 50 Mm CAPS [pH 10]), containing 1 Mm CaCl₂.

Results:

Preliminary findings have demonstrated strong inhibitory effects of lowconcentrations of the herbal extracts on a cocktail of proteases(containing high concentrations of matrix metalloproteinases 2, 3, 8 and9). These results demonstrate a direct inhibitory effect of low doses ofherbal extracts on common metalloproteinases.

Representative results are shown in the gelatin zymogram depicted inFIG. 2, in which active proteases are indicated as white bands on a darkbackground. Line 1: 50 ng active metalloproteases are clearlydetectable. Line 2 demonstrates definitive inhibition of the samemetalloprotease cocktail present in line 1 by a 1/50 dilution of theaforementioned herbal composition.

Example 6 In Vivo Treatment of Gingival Inflammation Using an HerbalComposition of the Invention: Effects on MMP Activity

This study forms part of a controlled double-blind matched-sample(sixteen patients), three part clinical trial of the use of a herbalcomposition of the invention in controlling gingival inflammation 1, 4and 7 days after placement of a transmucosal adhesive patch containing acomposition containing Echinacea purpurea, Sambucus nigra and Centellaasiatica (prepared as described in Example 2, hereinabove).

In the control subjects, a placebo treatment comprising a transmucosaladhesive patch containing food color was used.

Gingival Tissue removed during periodontal surgery was immediatelyplaced on ice and subsequently frozen and stored at −80 degrees C.,prior to performing matrix metalloproteinase (MMP) activity analysisthereon, as described hereinabove in Example 5. The gingival samples areprepared for this analysis by homogenizing the thawed tissue in PBS andcentrifuging [10000 g×10 min].

The preliminary results obtained (data not shown) demonstrate that bandswere found in the areas consistent with MMP 2, 9 which have beenidentified as proteases associated with periodontal disease. Tissuesamples taken from the experimental sites showed no protease activity,indicating complete inhibition by the herbal composition of theinvention.

Example 7 In Vivo Effect of an Herbal Composition of the Invention onLocalized Irritation Following an Insect Bite

A subject having a painful insect bite on the skin overlying the upperarm was treated for a period of 24 hours with an adhesive patchcomprising a composition containing Echinacea purpurea, Sambucus nigraand Centella asiatica (prepared as described in Example 2, hereinabove).The insect bite-associated pain experienced by the patient was recordedand expressed on a visual analog scale, as depicted in FIG. 3. Theclinical correlates of the pain index values used in this scale are asfollows: 0=asymptomatic; 50=requires medication; 100=extreme localizeddiscomfort. The highest recorded pain index reported in this study was38.

It may be seen from these results that almost instantaneous relief ofthe localized irritation was obtained.

Formulations

The following formulations comprising herbal compositions of theinvention are given for purposes of illustration and exemplificationonly, and are not intended to limit the scope of the invention in anyway. Thus, both the concentration of active ingredient within eachformulation may be changed without removing said formulation from thescope of the invention. Similarly, other formulations comprising theherbal compositions claimed herein that contain different carriers,diluents, excipients, colorings, flavorings and other additives arestill to be considered to be within the scope of the present invention.

The term “Active ingredient” used in the following formulation tablesrefers to any combination of herbal extracts that are within the scopeof the invention. The weight percentage of the active ingredient iscalculated in terms of the dry weight of the herbal composition.Representative examples of such combinations are:

A) composition comprising Echinacea purpurea, Hypericum perforatum,Commiphora molmol, Uncaria tomentosa and Sambucus nigra in a weightratio of 4:4:4:5:3.

B) composition comprising Echinacea purpurea, Sambucus nigra andCentella asiatica in a weight ratio of 15:70:15.

Formulation 1 Mouthwash

Ingredient % by weight Active ingredient 0.15 Glycerin, U.S.P 10.000Ethanol, 190-proof, U.S.P 7.500 Flavor 0.040 Polyoxythylene (20) 0.200sorbitan monoisostearate Sodium saccharin, N.P. 0.050 Boric acid, U.S.P0.075 FD&C Green (1% solution) 0.045 Distilled water balance

Formulation 2 Lozenge

Ingredient % by weight Active ingredient 0.25 Sorbitol 17.5 Mannitol17.5 Starch 13.6 Sugar substitute 1.2 Flavor 11.7 Color 0.1 Corn syrupBalance

Formulation 3 Chewing Gum

Ingredient % by weight Active ingredient 0.25 Gum base 30.00 (30 partsEastergum, 45 parts Coumarone resin, 15 parts dry latex, 10 partsParaffin wax) Sugar 50.00 Corn syrup 18.00 Citric acid 1.00 Flavorbalance

Formulation 4 Toothpaste

Ingredient % by weight Active ingredient 0.5 Sorbitol 33.00 Saccharin0.46 Silica 22.00 NaF 0.243 Glycerin 9.00 NaOH (50%) 0.20 Carbopol 0.20Keltrol 0.60 TiO₂ 0.50 Sodium alkyl sulphate 4.00 (28% solution) PEG 63.00 FD&C Blue# 1 (1% solution) 0.05 Flavor 1.1 Water Balance

While specific embodiments of the invention have been described for thepurpose of illustration, it will be understood that the invention may becarried out in practice by skilled persons with many modifications,variations and adaptations, without departing from its spirit orexceeding the scope of the claims.

What is claimed is:
 1. A therapeutic composition for treating oral/oranal mucosal diseases comprising effective amounts of extracts from thespecies Echinacea purpurea, Sambucus nigra and Centella asiatica,wherein the composition reduces inflammation, and wherein the amount ofeach of the extracts is in an amount that, if added to a correspondingcomposition of the other two extracts, provides an increased effect on amarker of gingival inflammation, and wherein the combination provides amore than additive increased effect on the marker.
 2. A therapeuticcomposition according to claim 1 for use in the treatment of diseases ofthe oral mucosa.
 3. A therapeutic composition according to claim 1 foruse in the treatment of diseases of the anal mucosa.
 4. A therapeuticcomposition according to claim 1, further comprising one or moreextracts of plants selected from the group consisting of Uncariatomentosa, Thymus vulgaris, Matricaria recutita, Salix alba, Calendulaofficinalis, Usnea barbata, Ligusticum porterii-osha, Gaultheriaprocumbens, Camellia sinensis, Vaccinium myrtillus, Melissa officinalis,Allium sativum, Camellia sinensis and Krameria triandra.
 5. Atherapeutic composition according to claim 4, wherein the disease of theoral mucosa to be treated is selected from the group consisting ofperiodontal disease, gingivitis, aphthous ulceration, mechanical trauma,thermal trauma, lichen planus, bullous pemphigoid, pemphigus vulgarisand dermatitis herpetiformis, angular chelitis and recurrent herpes. 6.A therapeutic composition according to claim 5, wherein the disease ofthe anal mucosa to be treated is selected from the group consisting ofanal fissures, hemorrhoids and non-specific irritation.
 7. A therapeuticcomposition according to claim 1, wherein the composition inhibits oneor more matrix metalloproteinases.
 8. A therapeutic compositionaccording to claim 7, wherein the one or more matrix metalloproteinasesto be inhibited are selected from the group consisting of matrixmetalloproteinases 1-9.
 9. A therapeutic composition according to claim8, wherein the matrix metalloproteinases that are inhibited are ofmatrix metalloproteinases of subclasses 1, 2, 8 and
 9. 10. A therapeuticcomposition according to claim 9, wherein the matrix metalloproteinasethat is inhibited is of subclass
 2. 11. A therapeutic compositionaccording to claim 10 for use in the treatment of a disease of the oralmucosa selected from the group consisting of periodontal disease andaphthous ulceration.
 12. A therapeutic composition according to claim 2,wherein the disease of the oral mucosa to be treated is selected fromthe group consisting of periodontal disease, gingivitis, aphthousulceration, mechanical trauma, thermal trauma, lichen planus, bullouspemphigoid, pemphigus vulgaris, dermatitis herpetiformis, angularchelitis and recurrent herpes.
 13. A therapeutic composition accordingto claim 3, wherein the disease of the anal mucosa to be treated isselected from the group consisting of anal fissures, hemorrhoids andnon-specific irritation.
 14. A therapeutic composition according toclaim 1, wherein the marker of gingival inflammation that is theproteolytic activity in gingival tissue of a matrix metalloproteinasesof subclasses 1 to
 9. 15. A therapeutic composition according to claim1, wherein the marker of gingival inflammation that is the proteolyticactivity in gingival tissue of a matrix metalloproteinases of subclasses2 or 9.